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    Salivary markers of systemic diseases ppt Salivary markers of systemic diseases ppt Presentation Transcript

    • SALIVARY MARKERS OF SYSTEMIC DISEASES
      BY
      Y.PRAGZNA(10T21S0113)
      UNDER THE GUIDANCE OF
      Mrs. M. SANDHYA M.PHARM
      ASSOCIATE PROFESSOR
    • CONTENTS:
      • Biomarkers
      • Saliva physiology
      • Advantages & disadvantages
      • Saliva collection, processing & storage
      • Diagnostic applications
      • Salivary proteome & transcriptome
      • New technologies of measurement
    • BIOMARKERS:
      Biomarkers are defined as cellular, biochemical and molecular characteristics by which normal/abnormal processes can be recognized and/or monitored.
    • SALIVARY MARKERS
      Non invasive diagnosis of disease and monitoring of general health
    • SALIVA PHYSIOLOGY:
      • Saliva is the mixed product of 3 major salivary glands.
      Parotid
      Sub mandibular
      Sub lingual
      • It is slightly acidic(pH 6-7).
      • Several physiological & pathological conditions modify saliva production quantitatively.
    • Functions of saliva
      PROTECTIVE
      • Lubrication
      • Anti-microbial
      • Mucosal integrity
      • Lavage/cleansing
      • Buffering
      • Remineralization
      FOOD & SPEECH RELATED
      • Food preparation
      • Digestion
      • Taste
      • Speech
      • Prevention of caries
    • SALIVA AS A DIAGNOSTIC FLUID
      ADVANTAGES
      • It is safe, non-invasive and simple.
      • No needles and No cringing.
      • Home testing.
      • It has an average protein concentration of 1.5-2 mg/ml.
      • Cost of shipping & Storage tend to be lower.
      Informative analytes are present in lower amounts in saliva than in serum.
      DISADVANTAGES
    • Whole saliva collection
      Ductal secretions collection
      Parotid secretions collection
      Sub mandibular & sub lingual secretions collection
      Saliva collection
    • PROCESSING & STORAGE:
      All procedures must be performed on ice:
      (1)Divide the saliva samples into multiple 330μL samples placed in cryotubes at -80ºc.
      Further processed for
      • Protein analysis: Add protease inhibitors like aprotinin(0.33µL), 3.3µL PMSF.
      • RNA analysis: Add 1.65µL of SUPERase inhibitor.
      (2)Invert gently to mix.
      (3)Store all fractions at -80ºc.
    • Salivary/Oral Fluid Biomarkers Possibilities of Use
      DNA Standard genotyping
      Genetic information of the hosting human body
      Bacterial infection (oral microbes in the mouth) Diagnosing carcinomas of head & neck
      Forensics
      mRNA Information on transcription rates
      RNA Viral/bacterial identification
      Proteins Genetic information
      Translational regulation
      Diagnosing periodontitis
      Detecting dental cavities
      Immunoglobulin Diagnosing viruses
      Metabolites Diagnosing periodontitis
    • DIAGNOSTIC APPLICATIONS OF SALIVA:
      • Drugs, hormones:
      • Intra cellular pathways
      Active & Passive diffusion
      • Extra cellular pathways
      Ultra filtration
      • Serum components:
      • Capillary barrier
      • Interstitial spaces
      • Cervicular fluid
      • Membrane of acinar
      & ductal cells
      • SJOGRENS SYNDROME:
      There are raised concentrations of NaCl, IgA, IgG, lactoferrin, inflammatory mediators such as PGE2, TXB2, Interleukin-6 etc.
      • CYSTIC FIBROSIS:
      Saliva of CF patients contain increased calcium & phosphate levels, urea & uric acid and abnormally elevated levels of PGE2.
      • DIABETES MELLITUS:
      It effects flow rate & composition of saliva. Salivary alterations includes higher levels of IgA, Salivary Peroxidase, glucose content, K, Salivary total protein & amylase.
      • ORAL MUCOSAL DISEASES:
      Nitric oxide levels were increased.
      • ONCOLOGY:
      P53 antibody can be detected in oral squamous cell carcinoma, increased levels of salivary kallikrein levels in oral malignant tumors, CA 15-3 in breast cancer. CVS:
      Determination of salivary amylase activity before 6hrs after cv surgery.
       KIDNEY DYSFUNCTION:
      Hypo salivation , changes in taste, ammonium smelling breath, oral mucosal pain.
      EPILEPSY:
      Gingival hypertrophy, IgA deficiency
    • VIRAL DISEASES:Acute hepatitis A & B were diagnosed based on the
      presence of IgM antibodies in saliva.HIV:Salivary IgA levels.BACTERIAL INFECTIONS:By monitoring the presence of antibodies to microorganisms found in saliva & oral cavity
      • AGAIN & AGAIN ABOUT TOBACCO:
      • Smoking causes decrease in salivary important antioxidant levels.
      • Salivary nicotine levels are considered to be the most reliable marker
    • Drug monitoring
      ANTI-ASTHMATICS:Theophylline
      ANTI EPILEPTICS: Carbamazapine, Diazepam, Ethosuximide, Lamotrigine, Topiramate, Phenytoin, Primidone, Valproic acid
      ANTI ARRYTHMIC/ANTI HYPERTENSIVES: Digoxin, Metoprolol, and Procainamide.
      ANTI MICROBIALS:Clarithromycin, Gentamycin, Isoniazid, Ofloxacin, Sulfanilamide
      ANTIPYRETICS: Acetaminophen, Antipyrine, Paracetamol
      ANTI NEOPLASTICS:Cisplatin, Methotrexate, Texol, Topotecan
      ANTI VIRAL: Quinine
      LITHIUM PSYCHOTROPIC:Lithium, Amitriptyline
      MISCELLANEOUS: Nicotine, Ethanol, Caffeine, Tolbutamide, Quinine
      ILLICIT DRUGS: Amphetamine, Barbiturates, Benzodiazepines, Opiods, Cocaine, Phencyclidine
    • HORMONE MONITORING:
      Steroids like cortisol, aldosterone, testosterone, insulin
      SIALOCHEMISTRY ANALYSIS:
      Early monitoring of an exposure to lead, cadmium, mercury because of higher salivary levels of these elements
    • CLASSIFICATION OF SALIVARY BIO MARKERS:
      • PROTEOMIC
      • GENOMIC
      • MICROBIAL
      • OTHERS
    • Diagnostic molecular targets
      SALIVARY PROTEOME:
      Use of ‘Divide & conquer’ strategy
      • Initial fractionation of proteins with separation techniques
      • Digestion with proteolytic enzyme, trypsin
      • Analysis by 1-D, 2-D LC-MS
      • Finally data is processed & submitted for database searching
    • SALIVARY PROTEOMIC ANALYSIS
    • SALIVARY TRANSCRIPTOME:
      • It includes mRNA molecules that cells use to convey the instructions carried by DNA for subsequent protein production.
      • At present, the main strategy for identification of salivary transcriptomic biomarkers is through microarray technology.
    • SALIVARY TRANSCRIPTOME:
      DNA MICROARRAY TECHNOLOGY
    • NEW TECHNOLOGIES FOR MEASURING SALIVARY BIOMARKERS:
      • POC DEVICE
      In this POC device, saliva(100-300μL) is placed into the salivary collection module, and then delivered into the Nano-biochip.
      • MICROBEAD ARRAY
      Micro pits within a silicon wafer are populated with a variety of chemically sensitized bead micro reactors.
    • MICRO BEAD ARRAY TECHNIQUE
      NANO-BIOCHIP
    • CONCLUSION:
      The components of saliva act as a “Mirror of the body’s health”,
      widespread use and growing acceptability of saliva as a
      diagnostic tool is helping individuals, researchers, health care
      professionals and community health programs to better detect
      and monitor diseases and to improve the general health of the
      public
    • REFERENCES:
      • Mandel ID. A contemporary view of salivary research. Crit Rev Oral Biol Med 1993; 4:599.
      • Raymond G, Schipper a, Erika Silletti a, Monique H, Vingerhoeds. Saliva as research material: biochemical, physiochemical, and practical aspects. Archives of Oral Biology 2007 ;( 52):1114-35.
      • Richard P, Jiri S, Jana V, Edgar F, Petr M, Jindrich P et al. Saliva as a diagnostic medium. Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub 2009; 153(2):103-10.
      • Eliaz Kaufman .The diagnostic applications of saliva. A review. Crit Rev Oral Biol Med 2002; 13(2):197-212.
      • www.med.kufauniv.com/ar/journals2010/res%20(1).pdf
      • www.jofs.in/files/Artical-11.pdf
      • www.linkgroup.hu/docs/pcs2/218-08CurrGen.pdf
      • www.wikipedia.com
       
    • THANK YOU
    • QUERIES???